Efficacy of five 'sporicidal' surface disinfectants against Clostridioides difficile spores in suspension tests and 4-field tests.

BACKGROUND: A sporicidal surface disinfection is recommended both for the outbreak and the endemic setting but a comparative evaluation on the efficacy of 'sporicidal' surface disinfectants using suspension tests and 4-field tests has not been performed. AIM: To determine the efficacy of five 'sporicidal' surface disinfectants (three ready-to-use wipes (A, B, E), two concentrates (C, D) based on peroxides or aldehydes against C. difficile spores. METHODS: The efficacy was determined under clean conditions using a suspension test and the 4-field test. Each test was performed in duplicate in two separate laboratories. Wipes were wrung to collect the solution for the suspension tests. RESULTS: Product A (peracetic acid; 5 min), product C (peracetic acid; 2% solution in 15 min or 1% solution in 30 min) and product D (peracetic acid; only 2% solution in 15 min) were effective with at least a 4 log10-reduction of C. difficile spores in suspension and on surfaces. Product B (hydrogen peroxide) was not effective in suspension (0.9 log10 after 15 min; 3.2 log10 after 1 h) and on surfaces (2.8 log10 after 15 and 60 min). Product E based on glutaraldehyde, (ethylendioxy)dimethanol and DDAC demonstrated 0.9 log10 after 4 h in suspension and 4.5 log10 after 4 h on surfaces. CONCLUSIONS: Not all surface disinfectants with a sporicidal claim were effective against C. difficile spores in standardized suspension tests and in the 4-field test. In clinical practice preference should be given to products that reliably pass the efficacy criteria of both types of tests.

Interlaboratory reproducibility of a test method following 4-field test methodology to evaluate the susceptibility of Clostridium difficile spores.

BACKGROUND: Sporicidal surface disinfection is recommended to control transmission of Clostridium difficile in healthcare facilities. EN 17126 provides a method to determine the sporicidal activity in suspension and has been approved as a European standard. In addition, a sporicidal surface test has been proposed. AIM: To determine the interlaboratory reproducibility of a test method for evaluating the susceptibility of a C. difficile spore preparation to a biocidal formulation following the 4-field test (EN 16615 methodology). METHODS: Nine laboratories participated. C. difficile NCTC 13366 spores were used. Glutaraldehyde (1% and 6%; 15 min) and peracetic acid (PAA; 0.01% and 0.04%; 15 min) were used to determine the spores' susceptibility in suspension in triplicate. FINDINGS: One-percent glutaraldehyde revealed a mean decimal log10 reduction of 1.03 with variable results in the nine laboratories (0.37-1.49) and a reproducibility of 0.38. The effect of 6% glutaraldehyde was stronger (mean: 2.05; range: 0.96-4.29; reproducibility: 0.86). PAA revealed similar results. An exemplary biocidal formulation based on 5% PAA was used at 0.5% (non-effective concentration) and 4% (effective concentration) to determine the sporicidal efficacy (4-field test) under clean conditions in triplicate with a contact time of 15 min. When used at 0.5% it demonstrated an overall log10 reduction of 2.68 (range: 2.35-3.57) and at 4% of 4.61 (range: 3.82-5.71). The residual contamination on the three primarily uncontaminated test fields was <50 cfu/25 cm2 in one out of nine laboratories (0.5%) and in seven out of nine laboratories (4%). CONCLUSION: The interlaboratory reproducibility seems to be robust.

Isopropanol at 60% and at 70% are effective against 'isopropanol-tolerant' Enterococcus faecium.

The bactericidal activity of isopropanol was determined against Enterococcus faecium ATCC 6057, ST 796 (isopropanol-tolerant strain) and Enterococcus hirae ATCC 10541 (EN 13727). Isopropanol at 60% and 70% were effective (≥5.38 log10-reduction) in 15 s against all strains but 23% isopropanol was not (<0.99 log10-reduction in ≤15 min). Isopropanol at 70% was tested against E. faecium in the four-field test. Eight millilitres was not effective enough in 1 min (<5 log10-reduction), whilst 16 mL was effective (≥5.85 log10-reduction). Healthcare workers can be reassured that 60% and 70% isopropanol with an appropriate volume are effective against E. faecium.

Verification of the efficiency of chemical disinfection and sanitation measures in in-building distribution systems.

Previous investigations of biofilms, generated in a silicone tube model have shown that the number of colony forming units (CFU) can reach 10(7)/cm(2), the total cell count (TCC) of microorganisms can be up to 10(8)cells/cm(2). The present study focuses on the situation in in-building distribution systems. Different chemical disinfectants were tested for their efficacy on drinking water biofilms in silicone tubes: free chlorine (electrochemically activated), chlorine dioxide, hydrogen peroxide (H(2)O(2)), silver, and fruit acids. With regard to the widely differing manufacturers' instructions for the usage of their disinfectants three different variations of the silicone tube model were developed to simulate practical use conditions. First the continuous treatment, second the intermittent treatment, third the efficacy of external disinfection treatment and the monitoring for possible biofilm formation with the Hygiene-Monitor. The working experience showed that it is important to know how to handle the individual disinfectants. Every active ingredient has its own optimal application concerning its concentration, exposure time, physical parameters like pH, temperature or redox potential. When used correctly all products tested were able to reduce the CFU to a value below the detection limit. Most of the active ingredients could not significantly reduce the TCC/cm(2), which means that viable microorganisms may still be present in the system. Thus the question arises what happened with these cells? In some cases SEM pictures of the biofilm matrix after a successful disinfection still showed biofilm residues. According to these results, no general correlation between CFU/cm(2), TCC/cm(2) and the visualised biofilm matrix on the silicone tube surface (SEM) could be demonstrated after a treatment with disinfectants.

Detection of Helicobacter pylori in biofilms by real-time PCR.

Helicobacter pylori is a cause of peptic ulcer disease and a causative agent of gastric cancer. Currently, a possible waterborne route of transmission or a possible survival in drinking water biofilms is discussed. H. pylori, like many other bacterial strains, has the ability to enter the viable but nonculturable state (vbnc) in case of unfavorable conditions. Therefore it is necessary to develop new analysis tools for vbnc bacteria. We established a fast and reliable method to detect H. pylori in drinking water biofilms by quantitative real-time PCR which makes it redundant to use difficult cultivation methods for nonculturable bacteria. With this method it was possible to identify water biofilms as a niche for H. pylori. The real-time PCR analysis targets the ureA subunit of the Helicobacter pylori urea gene which showed high specificity and sensitivity. The quantitative real-time PCR was used to detect H. pylori in biofilms of different age, unspiked and spiked with predetermined levels of cells. The drinking water biofilms were generated in a silicone-tube model. The DNA-sequences for probe and primers showed no cross-homologies to other related bacteria and it was possible to detect less than 10 genomic units of H. pylori. This novel method is a useful tool for a fast screening of drinking water biofilms for H. pylori. The results suggest that drinking water biofilms may act as a reservoir for H. pylori which raises new concerns about the role of biofilms as vectors for pathogens like Helicobacter pylori.